环状 RNA circ_0026218 通过 miR-338-3p/SIRT6 轴抑制动脉粥样硬化进展。
Circular RNA circ_0026218 Suppressed Atherosclerosis Progression via miR-338-3p/SIRT6 Axis.
机构信息
Heart Center, Guilin People's Hospital, Guilin, 541002 Guangxi, China.
出版信息
Biomed Res Int. 2023 Jan 24;2023:5647758. doi: 10.1155/2023/5647758. eCollection 2023.
BACKGROUND
Multiple circular RNAs (circRNAs) are implicated in atherosclerosis (AS) pathogenesis. In fact, how circRNA 0026218 (circ_0026218) functions in AS remains unknown, and thus the functions and mechanisms of circ_0026218 in the injury of vascular endothelial cells are to be investigated.
METHODS
Microarray analysis was employed to screen out differentially expressed circRNAs in AS. A cell model was mimicked by treating Human umbilical vein endothelial cells (HUVECs) with oxidized low-density lipoprotein (ox-LDL). circ_0026218, microRNA-338-3p (miR-338-3p) and silent information regulator 6 (SIRT6) expressions in HUVECs with ox-LDL treatment were probed by qRT-PCR. The cell proliferative capabilities were exposed by CCK-8 assay. The contents of interleukin 6 (IL-6), interleukin 1 (IL-1), and tumor necrosis factor (TNF-) were measured by ELISA. Oxidative stress kits were utilized to detect the levels of reactive oxygen species (ROS), superoxide dismutase (SOD), and malondialdehyde (MDA). Flow cytometry was adopted to analyze the level of apoptosis of HUVECs. Dual-luciferase reporter gene assay and RIP assay were leveraged to expose the interplay between miR-338-3p and circ_0026218 or SIRT6 3'-UTR, respectively. In addition, the impacts of circ_0026216 and miR-338-3p on SIRT6 protein expressions were subjected to Western blot.
RESULTS
circ_0026218 was greatly depleted in ox-LDL-stimulated HUVECs. circ_0026218 overexpression promoted viability of HUVECs and inhibited inflammatory response, oxidative stress, and apoptosis. circ_0026218 could adsorb miR-338-3p and positively modulated SIRT6 expressions via sponging miR-338-3p. Upregulation of this miRNA reversed the influence of circ_0026218 overexpression on ox-LDL-caused injury and apoptosis of HUVECs.
CONCLUSION
Collectively, circ_0026218 upregulates SIRT6 expression through decoying miR-338-3p, thereby inhibiting ox-LDL-initiated injury of HUVECs. circ_0026218 is involved in the pathogenesis of AS.
背景
多种环状 RNA（circRNA）被认为与动脉粥样硬化（AS）的发病机制有关。事实上，circRNA0026218（circ_0026218）在 AS 中的作用仍不清楚，因此需要研究 circ_0026218 在血管内皮细胞损伤中的功能和机制。
方法
采用微阵列分析筛选出 AS 中差异表达的 circRNA。用氧化型低密度脂蛋白（ox-LDL）处理人脐静脉内皮细胞（HUVEC）模拟细胞模型。用 qRT-PCR 检测 ox-LDL 处理的 HUVEC 中 circ_0026218、microRNA-338-3p（miR-338-3p）和沉默信息调节因子 6（SIRT6）的表达。用 CCK-8 法检测细胞增殖能力。用 ELISA 法测定白细胞介素 6（IL-6）、白细胞介素 1（IL-1）和肿瘤坏死因子（TNF-）的含量。用氧化应激试剂盒检测活性氧（ROS）、超氧化物歧化酶（SOD）和丙二醛（MDA）的水平。用流式细胞术分析 HUVEC 凋亡水平。分别利用双荧光素酶报告基因检测和 RIP 实验验证 miR-338-3p 与 circ_0026218 或 SIRT6 3'-UTR 的相互作用。此外，Western blot 检测 circ_0026216 和 miR-338-3p 对 SIRT6 蛋白表达的影响。
结果
ox-LDL 刺激的 HUVEC 中 circ_0026218 明显减少。circ_0026218 的过表达促进了 HUVEC 的活力，并抑制了炎症反应、氧化应激和凋亡。circ_0026218 可以吸附 miR-338-3p，并通过海绵吸附 miR-338-3p 正向调节 SIRT6 的表达。该 miRNA 的上调逆转了 circ_0026218 过表达对 ox-LDL 引起的 HUVEC 损伤和凋亡的影响。
结论
综上所述，circ_0026218 通过诱饵 miR-338-3p 上调 SIRT6 表达，从而抑制 ox-LDL 引发的 HUVEC 损伤。circ_0026218 参与 AS 的发病机制。
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